Competition based-detection assays

摘要

Disclosed herein are methods and kits which are useful for detecting presence of an enzyme and the relative amount of glycan associated with the enzyme in a test sample based upon the enzyme's ability to competitively inhibit the binding of a ligand in such test sample. The present invention provides the ability to evaluate cell culture conditions and optimize the desired glycoform content of recombinantly prepared enzymes.

主权项

1. A method of detecting a relative amount of glycan associated with an enzyme in a test sample, the method comprising: (a) contacting the test sample with at least one capture agent under conditions appropriate for binding of glycosylated enzyme in the test sample to the capture agent, wherein if the glycosylated enzyme is present in the test sample a bound enzyme is formed; (b) separating the bound enzyme from the test sample; c) contacting the bound enzyme with a ligand for the capture agent and (d) detecting the extent to which the bound enzyme inhibits binding of the ligand to the capture agent, wherein the extent to which the bound enzyme inhibits binding of the ligand to the capture agent is indicative of the relative amount of glycan associated with the enzyme in the test sample; wherein the capture agent comprises a biotinylated lectin.